NOTCH3-R544C mutation drives endothelial dysfunction through inflammation, migration impairment, and lipid...

Area of Science:

• Neuroscience
• Vascular Biology
• Genetics

Background:

• Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is linked to NOTCH3 gene mutations.
• Research has focused on smooth muscle cells, leaving brain microvascular endothelial cell (BMEC) roles unclear.

Purpose of the Study:

• Investigate the impact of the NOTCH3-R544C mutation on BMEC function.
• Elucidate mechanisms of endothelial dysfunction in CADASIL.

Main Methods:

• Utilized CADASIL transgenic mice and EC models with NOTCH3-R544C.
• Assessed endothelial function via immunofluorescence, RNA-seq, PPI network mapping, and lipid/cellular assays.

Main Results:

• NOTCH3 extracellular domain (NOTCH3ECD) deposition and reduced microvascular density observed in CADASIL mice.
• R544C mutant cells showed NOTCH3ECD accumulation, gene dysregulation (inflammation, migration), and a CXCL10-centered axis.
• R544C cells exhibited increased inflammation, cholesterol accumulation, reduced viability, and heightened sensitivity to stimuli.

Conclusions:

• The NOTCH3-R544C mutation impairs BMEC inflammatory regulation, migration, and lipid homeostasis.
• This disruption leads to endothelial dysfunction and reveals a mechanism of endothelial injury in CADASIL.